<Abstract>Objective: The aim of the study was to determine the level of bacterial contamination of door handles/knobs of toilets and bathrooms at Daeyang Luke Hospital.Methods: Fomite can be described as a non-living object capable of carrying infectious organism and it is considered as one of the major source of spread of hospital acquired infection. In this study, door handles/knobs were swabbed with sterile swab stick moistened with sterile water. The content was evenly transferred to nutrient agar petric dish and then incubated for 24 hours at 37˚C. The study was done between the months of May to August 2016.Results: The results of the study showed that during the period of study there was 41.6% bacterial contamination with the female medical and surgical ward toilets being more contaminated as compared to staff toilets. Private room bathrooms were more contaminated during the period of study as compared to the staff room bathrooms that were less contaminated. Among the bacterial contaminant isolated and identified, Staphylococcus aureus had the highest prevalence followed by Escherichia coli, Pseudomonas aeruginosa, Citrobacter species and Klebsiella pneumoniae respectively.Conclusions: Therefore, from the study, it can be concluded that general public needs observe hand hygiene and use of quality disinfection procedures in order to reduce hospital acquired infection.</Abstract>